Priming com INF-Ƴ e TNF-α aumenta potencial imunomodulador de células tronco mesenquimais caninas in vitro
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Data
2023-03-23
Autores
Chimenes, Natielly Dias
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A meningoencefalite de origem desconhecida (MUO) é uma enfermidade neuroimune em cães, com grande potencial de se tornar um modelo animal natural para a esclerose múltipla. O priming in vitro das células tronco mesenquimais derivadas de tecido adiposo canino (cAd-MSCs) é uma estratégia para potencializar a ação imunomodulatória e imunossupressora das cAd-MSCs. O objetivo deste trabalho foi avaliar o priming com IFN-Ƴ e TNF-α, LCR e soro de cães com MUO nas cAd-MSCs. Para tanto, foi realizado o priming in vitro por 72 horas das cAd-MSCs (P3-P4) em seis grupos: controle (C), com IFN-Ƴ em associação com TNF-α (IFN-Ƴ+ TNF-α), soro controle (C-S), soro de cães com MUO (MUO-S), LCR controle (C-LCR) e LCR de cães com MUO (MUO-LCR) e avaliada a expressão gênica para BDNF, GNDF, HGF, IDO, PTGE2, IL-10, IFN-Ƴ e TNF-α por qPCR e quantificação proteica de GM-CSF, IL-10, IL-2, IL-8 e MCP-1 por ensaio multiplexado. Comparando todos os grupos com o grupo C, o IFN-Ƴ+ TNF-α apresentou aumento de HGF, IDO e TNF-α e IL-8 (p= <0,001), nos grupos C-LCR e MUO-LCR houve aumento de IFN-Ƴ e TNF-α e redução de BDNF e GNDF (p= <0,001), os grupos C-S e MUO-S apresentaram redução na quantificação proteica de IL-10 (p= <0,001), e o grupo C-S teve redução de BDNF e GNDF (p= <0,001) e o grupo MUO-S teve redução de HGF (p= <0,001). Diante dos resultados, a técnica priming IFN-Ƴ + TNF-α demonstrou maior potencial de perfil imunomodulatório com tendência anti-inflamatória em comparação com os demais grupos.
Meningoencephalitis of unknown origin (MUO) is a neuroimmune disease affects dogs, with great potential to become a natural experimental model for multiple sclerosis. In vitro priming of mesenchymal stem cells derived from canine adipose tissue (cAd-MSCs) is a strategy to enhances the immunomodulatory and immunosuppressive action of cAd-MSCs. The aim of this work is to evaluate the priming with IFN-γ and TNF-α, CSF and serum from dogs with MUO on cAd-MSCs. Therefore, in vitro priming of cAd-MSCs was performed in six groups: control (C), with IFN-α in association with TNF-α (IFN-α + TNF-α), control serum (C-P), serum from dogs with MUO (MUO-P), CSF control (C-CSF) and CSF from dogs with MUO (MUO-CSF) and evaluated gene expression for BDNF, GNDF, HGF, IDO, PTGE2, IL-10, IFN- Ƴ and TNF-α by qPCR and protein expression of GM-CSF, IL-10, IL-2, IL-8 and MCP-1 by multiplex assay. In comparison with group C, the group with IFN-γ+ TNF-α showed an increase in HGF, IDO and TNF-α and IL-8 (p= <0.001), in the C-CSF and MUO-CSF groups there was an increase in IFN -Ƴ and TNF-α and a reduction in BDNF and GNDF (p= <0.001), the C-S and MUO-S groups showed a reduction in IL-10 (p= <0.001), and the C-S group had a reduction in BDNF and GNDF ( p= <0.001) and the MUO-S group had reduced HGF (p= <0.001). In view of the results, the IFN-γ + TNF-α priming technique demonstrated a greater potencial for immunomodulatory profile with an anti-inflmmatory tendency compared to the other groups.
Meningoencephalitis of unknown origin (MUO) is a neuroimmune disease affects dogs, with great potential to become a natural experimental model for multiple sclerosis. In vitro priming of mesenchymal stem cells derived from canine adipose tissue (cAd-MSCs) is a strategy to enhances the immunomodulatory and immunosuppressive action of cAd-MSCs. The aim of this work is to evaluate the priming with IFN-γ and TNF-α, CSF and serum from dogs with MUO on cAd-MSCs. Therefore, in vitro priming of cAd-MSCs was performed in six groups: control (C), with IFN-α in association with TNF-α (IFN-α + TNF-α), control serum (C-P), serum from dogs with MUO (MUO-P), CSF control (C-CSF) and CSF from dogs with MUO (MUO-CSF) and evaluated gene expression for BDNF, GNDF, HGF, IDO, PTGE2, IL-10, IFN- Ƴ and TNF-α by qPCR and protein expression of GM-CSF, IL-10, IL-2, IL-8 and MCP-1 by multiplex assay. In comparison with group C, the group with IFN-γ+ TNF-α showed an increase in HGF, IDO and TNF-α and IL-8 (p= <0.001), in the C-CSF and MUO-CSF groups there was an increase in IFN -Ƴ and TNF-α and a reduction in BDNF and GNDF (p= <0.001), the C-S and MUO-S groups showed a reduction in IL-10 (p= <0.001), and the C-S group had a reduction in BDNF and GNDF ( p= <0.001) and the MUO-S group had reduced HGF (p= <0.001). In view of the results, the IFN-γ + TNF-α priming technique demonstrated a greater potencial for immunomodulatory profile with an anti-inflmmatory tendency compared to the other groups.
Descrição
Palavras-chave
Neuroimunes, Citocinas, Meningoencefalite, Esclerose múltipla